背景:犬炎性乳腺癌(CIMC)的转移发生率很高,高杀伤力,预后不良,这需要新的佐剂。川芎嗪-大黄酸衍生物(TRD)已被证明具有抗肿瘤活性,这是CIMC的一个潜在研究方向。
目的:本研究评估了TRD在体外和体内对CIMC的疗效,为活性化合物在中药中的应用提供了可能。
方法:体外,用CCK-8测量TRD细胞毒性。流式细胞术和透射电镜检测细胞周期,细胞死亡,和线粒体的变化。伤口愈合试验,细胞侵袭试验,扫描电子显微镜观察细胞迁移和侵袭的抑制作用。通过RT-qPCR和Westernblot检测表达变化。在体内,将肺转移模型随机分为对照组,低剂量TRD,高剂量TRD,积极的团体。每组每天口服一次,共18天,并拍摄体内成像照片。
结果:TRD在CHMp和MDCK中的IC50分别为42.59和79.37μM,分别。TRD经由过程线粒体毁伤介导细胞凋亡,并经由过程下调细胞周期蛋白B1惹起S和G2/M期阻滞。此外,TRD通过下调钙黏着蛋白减少丝足病并抑制细胞迁移。InCIMC肺转移模型,TRD能有效抑制肺部肿瘤生长(P<0.001),无明显毒性。
结论:TRD具有抑制CIMC肺转移的潜在活性,具有多靶点和低毒性。
BACKGROUND: Canine inflammatory mammary carcinoma (CIMC) has a high incidence of metastasis, high lethality, and poor prognosis, which needs novel adjuvant agents. Tetramethylpyrazine-Rhein Derivative (TRD) has been shown to have antitumor activity, which is a potential research direction for CIMC.
OBJECTIVE: This study evaluated the efficacy of TRD on CIMC in vitro and in vivo, and provided possibilities for the application of active compounds in traditional Chinese medicine.
METHODS: In vitro, TRD cytotoxicity was measured with CCK-8. Flow cytometry and transmission electron microscope were used to detect the cell cycle, cell death, and changes in mitochondria. Wound-healing assay, cell invasion assay, and scanning electron microscope were used to evaluate the suppression of cell migration and invasion. Expression changes were detected by RT-qPCR and western blot assay. In vivo, the lung metastasis models were randomly divided into control, low-dose TRD, high-dose TRD, and positive groups. Each group was administered orally once a day for 18 days and took in vivo imaging photos.
RESULTS: The IC50 of TRD in CHMp and MDCK were 42.59 and 79.37 μM, respectively. TRD mediated cell apoptosis by mitochondrial damage and caused S and G2/M phase arrest by downregulating cyclin B1. Moreover, TRD reduced filopodia and inhibited cell migration by downregulating cadherins. In CIMC lung metastasis models, TRD could effectively inhibit tumor growth (P < 0.001) in the lungs without significant toxicity.
CONCLUSIONS: TRD showed potential activity to inhibit CIMC lung metastasis with multi-target and low toxicity.